Study on the mechanism of miRNAs on liver injury in the condition of Protoscocephalus alveolarus transhepatic portal vein infection.

OBJECTIVE: To explore the role of miRNA in liver damage caused by Echinococcus multilocularis infection. METHODS: Six female C57BL mice were randomly divided into two groups, the control group and the infection group. Mice in the control group were injected with 100 µL PBS through the hepatic portal vein, and mice in the infection group were infected with E. multilocularis via the hepatic portal vein to establish a mouse model of infection. Small RNA sequencing was performed for detecting the expression of miRNAs in the liver of mice infected with 2000 E. multilocularis after 3 months of infection, screen out miRNAs related to liver damage, and verify by RT-PCR. RESULTS: Seventy-one differentially expressed miRNAs were found in the liver in comparison with control, and a total of 36 mouse miRNAs with |FC| >0.585 were screened out, respectively. In addition, Targetscan (V5.0) and miRanda (v3.3a) software were used to predict differential miRNAs target genes and functional enrichment of target genes. Functional annotation showed that "cytokine-cytokine interaction," "positive regulation of cytokine production," "inflammatory response," and "leukocyte activation" were enriched in the liver of E. multilocularis-infected mice. Moreover, the pathways "human cytomegalovirus infection," "cysteine and methionine metabolism," "Notch signaling pathway," and "ferroptosis" were involved in liver disease. Furthermore, four miRNAs (mmu-miR-30e-3p, mmu-miR-203-3p, mmu-miR-125b-5p, and mmu-miR-30c-2-3p) related to liver injury were screened and verified. CONCLUSION: This study revealed that the expression profiling of miRNAs in the livers was changed after E. multilocularis infection, and improved our understanding of the transcriptomic landscape of hepatic echinococcosis in mice.

An ultrasound-based referential of body height-adjusted normal liver organometry in school children from Bokito in rural Cameroon.

The grading system for ultrasonographic assessment of Schistosoma mansoni morbidity is crucial for evaluation of control programs. This requires prior definition of normal liver organometric ranges in the population from the endemic area. A cross-sectional study was conducted in a S. mansoni endemic area in rural Cameroon. 1002 Participants were screened and 234 of them, free from all common liver-affecting diseases in the area (schistosomiasis, malaria, hepatitis B and C) and with no ultrasonographic signs of liver disease were selected and their liver parameters measured by ultrasonography. All statistics were considered significant for p-values < 0.05. Normal dimensions of livers lobe sizes, portal vein wall thickness and portal vein diameters are reported. The liver organometric data are presented for the entire study population as a whole and separately for males and females as prediction plots, with observed values and fitted regression line with 95% confidence. Reference ranges for liver parameters (size, portal vein thickness and diameter) adjusted for body height established in the current study are novel for Cameroon. The prediction plots generated should improve the accuracy of the assessment of liver morbidity by ultrasonography in the region.

Venous thrombosis and segmental hypoperfusion in amebic liver abscess: MDCT demonstration and its implications.

PURPOSE: To report venous thrombosis and associated perfusion defect in amebic liver abscess (ALA) using MDCT. METHOD: MDCT images of 62 patients with ALA were reviewed for venous thrombosis and associated perfusion abnormalities. RESULT: The study found 43 (69%) patients with venous thrombosis: portal vein thrombosis (PVT) occurred in 39, hepatic vein thrombosis (HVT) in 37 and inferior vena cava (IVC) thrombosis in 4. Combined PVT and HVT occurred in 33 (77%) patients. The portal vein thrombi remained localized in subsegmental branches in 25 patients and extended to segmental branches in 14. The hepatic vein thrombi were confined to peripheral branches in 18 patients; they progressed to the main trunk in 19 and to the IVC in 4. A wedge-shaped hypoattenuating zone suggesting ischemia was identified in 33 (77%) patients in portal phase: 31 had combined PVT and HVT, 2 had HVT alone, but none had PVT alone. It occurred significantly more often with combined PVT and HVT than HVT alone (p = 0.05). Arterial phase enhancement occurred in 2 of 13 patients with multiphasic CT. All patients were symptomatic despite medical therapy and therefore required percutaneous drainage. About half of the patients were identified with ruptured abscesses. Segmental atrophy was observed in seven of nine patients who underwent follow-up CT. CONCLUSION: Combined PVT and HVT commonly occur with ALA and often manifests as segmental hypoperfusion in portal venous phase, indicating ischemia. The detection of such events by CT may be indicative of severe disease that requires aggressive management involving percutaneous drainage.

Transhepatic-intrahepatic branches of the portal vein catheterization for ex vivo liver resection and autotransplantation: Two case reports of novel approach to perfuse the liver.

RATIONALE: There has been increased use of ex vivo liver resection and autotransplantation (ERAT) for treatment of end-stage hepatic alveolar echinococcosis (HAE). Rapid perfusion of the autograft in bench resection is always required to reduce the warm ischemia time (WIT) and to protect the function of the remnant liver. Nevertheless, the severe invasion of the portal hepatis sometimes makes it impossible to find a usable inflow rapidly and the process of perfusion could be delayed. PATIENT CONCERNS: Two patients diagnosed with end-stage HAE combined with severe portal hepatis invasion were selected to undergo ERAT at our center. DIAGNOSIS: Besides the large HAE lesions, the CT imaging of patient 1 showed that part of the intra- and extrahepatic portal vein (PV) had disappeared. Patient 2 had severe invasion of both of the right and left branches of the PV. INTERVENTIONS: We introduced a new approach for perfusing the liver in ERAT using transhepatic-intrahepatic branches of the PV catheterization. Afterward, ERAT was successfully performed. OUTCOMES: For patient 1, the WIT was 2 minutes and the cold ischemia time (CIT) was 296 minutes. For patient 2, the WIT was 2 minutes and the CIT was 374 minutes. Patient 1 suffered stenosis of the common bile duct on postoperative day 14, and patient 2 recovered uneventfully. Both of the 2 patients were discharged from the hospital with normal laboratory values on postoperative day 31 and 15, respectively. The laboratory values for both patients at recent follow-up were normal. LESSONS: Transhepatic-intrahepatic branches of the PV catheterization is useful for decreasing WIT and facilitating the management of ERAT. It is a useful technical variant that could be used in ERAT for treating patients with severe portal hepatis invasion.

Hydatid disease of the liver with portal vein invasion mimicking portal vein thrombosis.

Hydatid cyst disease is a zoonosis caused by the parasite Echinococcus. It may infest any organ of the body, but it most frequently involves the liver, lungs, and nervous system. Portal vein involvement by hydatid cyst disease is extremely rare with only five cases published in the English literature to our knowledge. We present the ultrasonography (US) and computed tomography (CT) findings of a 77-year-old male with hydatid disease of the liver with portal vein invasion mimicking portal vein thrombosis. Colour Doppler US confirmed the lack of blood flow within the portal vein and stigmata of cavernomatosis. CT clearly demonstrated a communication between the multiloculated lesion and the portal vein and the multiple daughter vesicles obstructing the portal vein. The consideration of this complication will make it possible to distinguish this entity from portal vein thrombosis and, thus, the management of the patients with hydatid cyst disease particulary in endemic regions.

Vascular infiltration-based surgical planning in treating end-stage hepatic alveolar echinococcosis with ex vivo liver resection and autotransplantation.

BACKGROUND: For end-stage hepatic alveolar echinococcosis, insufficient guidance is available regarding surgical treatment, especially for ex vivo liver resection combined with autotransplantation. The indications for this complex surgery require further discussion. METHOD: We reviewed 50 cases of patients who underwent ex vivo liver resection combined with autotransplantation from January 2014 to February 2018. A newly developed classification was used to describe vascular infiltration in all patients, who were divided into four groups based on anatomic lesion features and surgical patterns. The surgical planning for ex vivo liver resection combined with autotransplantation is then thoroughly discussed according to the gathered information. RESULTS: In all patients, the length of the operation and the anhepatic phase were 735 minutes (range, 540-1,170 minutes) and 309 minutes (range, 122-480 minutes), respectively. The median remnant liver volume-to-standard liver volume ratio was 0.58 (range, 0.32-1.11). The rate of complications classified as Clavien-Dindo grade III or higher was 22% (11/50). A total of 3 postoperative deaths occurred. We identified 4 types with distinguished lesion anatomic features. Type I patients required more frequent unconventional reconstruction of the portal vein and bile duct than the other patients. Of the 6 type IV patients, 4 required modification of the surgical protocol according to intraoperative findings. CONCLUSION: Vascular infiltration-based classification could improve the anatomic comprehension and, thus, facilitate surgical planning for ex vivo liver resection combined with autotransplantation. Through cautious evaluation of operability, liver function, and residual liver volume, together with delicate operative techniques and careful postoperative management, ex vivo liver resection combined with autotransplantation can achieve good results in the treatment of end-stage hepatic alveolar echinococcosis.

Phylogenetic Placement of a Schistosome from an Unusual Marine Snail Host, the False Limpet (Siphonaria lessoni) and Gulls (Larus dominicanus) from Argentina with a Brief Review of Marine Schistosomes from Snails.

In the blood fluke family Schistosomatidae, marine snails are well known as intermediate hosts. Eight families of marine snails have thus far been reported to host schistosomes across the world, most of which have been implicated in human cercarial dermatitis (HCD) outbreaks. As part of our larger effort to define the species diversity and biology of schistosomes in Argentina, in particular their role in causing HCD, we searched in the marine pulmonate snail (Siphonaria lessoni) for a schistosome species described previously from S. lessoni from southern Argentina. Additionally, gulls (Larus dominicanus) collected from a different project locality (inland) were examined, because they are known to spend time in the intertidal regions. Schistosome sporocysts were found in S. lessoni, and a small worm fragment was retrieved from a gull. Molecular phylogenies for 28S, ITS1-5.8S-ITS2, and cox1 genes revealed that the specimens from the gull and S. lessoni grouped closely together, suggesting they are conspecifics. Also, ITS1-5.8S-ITS2 sequences suggested one of the schistosomes from S. lessoni and a schistosome from a South African penguin were also conspecifics. Further study is needed to verify if these specimens comprise a distinct marine clade within the larger avian schistosome clade that is comprised mostly of species using freshwater snail hosts. Thus far, it appears this group of marine schistosomes may be more likely found in the southern hemisphere. It is unclear if the observed distribution pattern of schistosomes in Siphonaria is a result of sampling bias and/or indicative of a specific bird-snail-schistosome association. It is clear they are sharply differentiated from the basal marine clade of avian schistosomes that includes Austrobilharzia.

Hepatectomy for patients with alveolar echinococcosis: Long-term follow-up observations of 144 cases.

BACKGROUND: Western China is a region in which alveolar echinococcosis (AE) is endemic. Few studies and comparisons have evaluated the outcomes of AE patients after hepatectomy, and no strategy has been defined for the treatment of AE patients with unresectable tumors. This study sought to assess the outcomes of AE patients after hepatectomy at a tertiary referral center. PATIENTS AND METHODS: We retrospectively analyzed data from 144 patients with hepatic AE who were treated via hepatectomy at our center between January 2004 and December 2015. Patients' overall survival (OS), progression-free survival (PFS), and risk factors were analyzed, and Kaplan-Meier survival curves were constructed. Patient age, year of initial treatment, PNM stage, and risk factors were entered as co-variates in a Cox regression modle that was used for analysis. RESULTS: Hepatectomy was performed in 144 patients diagnosed with hepatic AE (84 complete resections and 60 reduction surgeries). In the complete resection group, the 5- and 10-year OS rates were both 97.6%, and the 5- and 10-year PFS rates were both 97.9%. In the reduction surgery group, the 5-, and 10-year OS rates were 89.7% and 73.4%, respectively, and the 5-, and 10-year PFS rates were 78.1% and 69.5%, respectively. Patients in the complete group had better OS prognoses and PFS than patients in the reduction surgery group (P = 0.018 and P = 0.001). Multivariate analysis indicated that curability and portal vein invasion are independent factors associated with PFS (P = 0.028 and P = 0.006). CONCLUSIONS: The most effective therapy for AE is complete resection. Reduction surgery does not appear to offer obvious advantages over benzimidazole therapy alone in the treatment of AE. Curability and portal vein invasion are independent prognostic factors for PFS in a multivariate analysis.

Portal vein thrombosis associated with protein C deficiency and elevated Factor VIII in hepatosplenic schistosomiasis.

Portal vein thrombosis is considered a vaso-occlusive process that can appear during the course of hepatosplenic Schistosoma mansoni, but may result from impaired portal blood flow or be associated with acquired or inherited thrombophilic factors. Here, we report the case of a 67-year-old woman who developed thrombocytopenia as a result of hypersplenism. Following the diagnosis of hepatosplenic schistosomiasis, portal vein thrombosis was detected by ultrasound examination, while haematological tests revealed low levels of protein C (43.3%) and high levels of factor VIII (183.1%). The pathogenesis of portal vein thrombosis remains unclear in some patients with S. mansoni. We recommend, therefore, that early clinical and haemostatic investigations are done to evaluate risk of portal vein thrombosis and hence avoid further complications.

Treatment and education reduce the severity of schistosomiasis periportal fibrosis.

INTRODUCTION: This study evaluates the factors associated with the development of severe periportal fibrosis in patients with Schistosoma mansoni. METHODS: A cross-sectional study was conducted from April to December 2012 involving 178 patients infected with S. mansoni who were treated in the Hospital das Clínicas of Pernambuco, Brazil. Information regarding risk factors was obtained using a questionnaire. Based on the patients' epidemiological history, clinical examination, and upper abdomen ultrasound evaluation, patients were divided into 2 groups: 137 with evidence of severe periportal fibrosis and 41 patients without fibrosis or with mild or moderate periportal fibrosis. Univariate and multivariate analyses were conducted using EpiInfo software version 3.5.5. RESULTS: Illiterate individuals (30.1%) and patients who had more frequent contact with contaminated water in towns in the Zona da Mata of Pernambuco (33.2%) were at greater risk for severe periportal fibrosis. Based on multivariate analysis, it was determined that an education level of up to 11 years of study and specific prior treatment for schistosomiasis were preventive factors for severe periportal fibrosis. CONCLUSIONS: The prevailing sites of the severe forms of periportal fibrosis are still within the Zona da Mata of Pernambuco, although there has been an expansion to urban areas and the state coast. Specific treatment and an increased level of education were identified as protective factors, indicating the need for implementing social, sanitary, and health education interventions aimed at schistosomiasis to combat the risk factors for this major public health problem.

Liposomal-praziquantel: efficacy against Schistosoma mansoni in a preclinical assay.

Currently, schistosomiasis mansoni is treated clinically with praziquantel (PZQ). Nevertheless, cases of tolerance and resistance to this drug have been reported, creating the need to develop new drugs or to improve existing drugs. Considering the small number of new drugs against Schistosoma mansoni, the design of nanotechnology-based drug delivery systems is an important strategy in combating this disease. The aim of this study was to evaluate the activity of PZQ containing liposome (lip.PZQ) on S. mansoni, BH strain. Mice were treated orally with different concentrations of PZQ and lip.PZQ 30 and 45 days following infection. The number of worms, recovered by perfusion of the hepatic portal system, and the number of eggs found in the intestine and liver were analysed. Parasite egg counts were also performed. The most active formulation for all parameters was 300mg/kg of lip.PZQ, since as it decreased the total number of worms by 68.8%, the number of eggs in the intestine by 79%, and the number of hepatic granulomas by 98.4% compared to untreated controls. In addition, this concentration decreased egg counts by 55.5%. The improved efficacy of the treatment with lip.PZQ, especially when administered 45 days following infection, compared with the positive-control group (untreated) and the groups that received free PZQ, can be explained by greater bioavailability in the host organism; the preferred target of lip.PZQ is the liver, and lip.PZQ is better absorbed by the tegument of S. mansoni, which has an affinity for phospholipids.

Treatment and education reduce the severity of schistosomiasis periportal fibrosis

Introduction This study evaluates the factors associated with the development of severe periportal fibrosis in patients with Schistosoma mansoni. Methods A cross-sectional study was conducted from April to December 2012 involving 178 patients infected with S. mansoni who were treated in the Hospital das Clínicas of Pernambuco, Brazil. Information regarding risk factors was obtained using a questionnaire. Based on the patients' epidemiological history, clinical examination, and upper abdomen ultrasound evaluation, patients were divided into 2 groups: 137 with evidence of severe periportal fibrosis and 41 patients without fibrosis or with mild or moderate periportal fibrosis. Univariate and multivariate analyses were conducted using EpiInfo software version 3.5.5. Results Illiterate individuals (30.1%) and patients who had more frequent contact with contaminated water in towns in the Zona da Mata of Pernambuco (33.2%) were at greater risk for severe periportal fibrosis. Based on multivariate analysis, it was determined that an education level of up to 11 years of study and specific prior treatment for schistosomiasis were preventive factors for severe periportal fibrosis. Conclusions The prevailing sites of the severe forms of periportal fibrosis are still within the Zona da Mata of Pernambuco, although there has been an expansion to urban areas and the state coast. Specific treatment and an increased level of education were identified as protective factors, indicating the need for implementing social, sanitary, and health education interventions aimed at schistosomiasis to combat the risk factors for this major public health problem. .

Proteomic identification of endothelial cell surface proteins isolated from the hepatic portal vein of mice infected with Schistosoma bovis.

Schistosomes are blood parasites adapted to their intravascular habitat that have evolved mechanisms to evade the immune and hemostatic responses of their hosts. It has been observed that the schistosome can regulate the endothelium function along the infection, which contributes to modulation of host defensive responses and parasite survival. The purpose of this work was the analysis of the changes induced by Schistosoma bovis adult worms in the proteome expressed by infected mice on the endothelial surface of their portal vein. With this aim, we have utilized a methodology that allows the purification, identification and relative quantification of endothelial cell surface proteins after their selective in vivo labeling with biotin. Trypsin digestion of the biotinylated proteins and subsequent liquid chromatography and tandem mass spectrometry analysis (LC-MS/MS) resulted in the identification of a total 127 non-redundant proteins. All these proteins have been classified according to their function and cellular location, and the differences between S. bovis-infected and non-infected mice in their endothelial surface proteomes have been analyzed. The present work provides the first data on the proteome of the endothelial surface of the portal vein, and identifies some of the changes induced in it after an infection by S. bovis.

Towards the establishment of a porcine model to study human amebiasis.

BACKGROUND: Entamoeba histolytica is an important parasite of the human intestine. Its life cycle is monoxenous with two stages: (i) the trophozoite, growing in the intestine and (ii) the cyst corresponding to the dissemination stage. The trophozoite in the intestine can live as a commensal leading to asymptomatic infection or as a tissue invasive form producing mucosal ulcers and liver abscesses. There is no animal model mimicking the whole disease cycle. Most of the biological information on E. histolytica has been obtained from trophozoite adapted to axenic culture. The reproduction of intestinal amebiasis in an animal model is difficult while for liver amebiasis there are well-described rodent models. During this study, we worked on the assessment of pigs as a new potential model to study amebiasis. METHODOLOGY/PRINCIPAL FINDINGS: We first co-cultured trophozoites of E. histolytica with porcine colonic fragments and observed a disruption of the mucosal architecture. Then, we showed that outbred pigs can be used to reproduce some lesions associated with human amebiasis. A detailed analysis was performed using a washed closed-jejunal loops model. In loops inoculated with virulent amebas a severe acute ulcerative jejunitis was observed with large hemorrhagic lesions 14 days post-inoculation associated with the presence of the trophozoites in the depth of the mucosa in two out four animals. Furthermore, typical large sized hepatic abscesses were observed in the liver of one animal 7 days post-injection in the portal vein and the liver parenchyma. CONCLUSIONS: The pig model could help with simultaneously studying intestinal and extraintestinal lesion development.

Parasitological and morphological study of Schistosoma mansoni and diabetes mellitus in mice.

Schistosomes are blood-dwelling flukes which are highly dependent on the host metabolism. The aim of this study was to investigate possible relationship between streptozotocin-induced diabetes and the outcome of acute murine schistosomiasis mansoni. Male and female SW mice were treated by a single intraperitoneally injected dose of streptozotocin (180 mg/kg). Seven days after induction, both control and diabetic animals were infected with 70 Schistosoma mansoni cercariae (BH strain). Diabetics and their controls were weighed 45 days after birth and for the last time prior to killing. Susceptibility to infection was evaluated twice a week by quantifying fecal egg excretion 7-9 weeks post-infection by the Kato-Katz' thick smear method. Mice were euthanized the day after the last fecal examination was performed. Adult worms were recovered from the portal system and mesenteric veins, whereas liver and intestine were removed for enumeration of egg load. No differences in worm length or in measurements of the reproductive organs, tegument, and suckers were detected. Also oviposition was unaffected as the total number of eggs per female worm from the liver, the small and the large intestine was the same in both groups. An oogram evaluation revealed a lower percentage of mature (23.0% vs. 40.7%) and a higher percentage of immature (69.1% vs. 51.7%) eggs in the small intestine of the diabetic mice. We suggest that principally a hampered egg passage through the intestine tissue caused this reduction and that probably both the eggs and the impaired host response play a role.

Expression, characterization and therapeutic efficacy of chimeric Fab of anti-idiotypic antibody NP30 against Schistosoma japonicum.

The murine monoclonal anti-idiotypic antibody NP30 is a promising therapeutic antibody against Schistosoma japonicum. However, the immunogenicity of murine NP30 limits its further study and application in humans. Here the chimeric Fab of NP30 (chFab-NP30) comprising the variable regions of murine NP30 and constant regions of human antibody was assembled. chFab-NP30 was expressed and purified as a soluble and functional protein. Administration of chFab-NP30 in vivo increased the survival rate, reduced egg burdens and ameliorated organ pathology of mice with acute schistosomiasis. Our study indicated that chFab-NP30 is a promising candidate to be used as a specific and efficient recombinant antibody against acute schistosomiasis japonica. Further studies on function mechanism of chFab-NP30 needs to be carried out in the future.

Potential effects of Cramoll 1,4 lectin on murine Schistosomiasis mansoni.

Cratylia mollis is a natural forage plant from the Northeast of Brazil. C. mollis seed lectin (Cramoll) containing molecular forms 1 and 4 (Cramoll 1,4) has shown anti-inflammatory and wound-healing activities. This work analyzed the effect of Cramoll 1,4 on experimental schistosomiasis in mice. Experimental groups (n=15/group) were composed of female albino Swiss mice, which were subcutaneously and caudally infected with Schistosoma mansoni (BH strain, 100 cercariae/mouse) and were treated with an intraperitoneal dose after infection as follows: (1) Cramoll 1,4 (50 mg kg(-1) single dose - after 40 days of infection), (2) Cramoll 1,4 (7 mg kg(-1) daily dose - for 7 days after infection) and control (untreated mice). Mice were sacrificed 8 weeks after infection and adult worms were recovered from the portal-hepatic system. Livers were fixed in 10% (v/v) formaldehyde/0.15M NaCl and tissue sections were processed for haematoxilin and Masson's trichrome stainings. Mice infected subcutaneously harboured no or very few worms and hence the effect of Cramoll 1,4 could not be assessed. Results (P≤0.05) were obtained with Cramoll 1,4 using the two treatments, with reduction of: egg excretion (79 and 80%), adult worm recovery (71 and 79%) and liver granulomas (40 and 73.5%) in relation to control. This study showed the potential anti-helminthic activity of Cramoll 1,4 when tested against Schistosomiasis mansoni infection in mice.